Development of enhanced primer sets for detection of Norovirus and Hepatitis A in food samples from Guayaquil (Ecuador) by reverse transcriptase-heminested PCR

Abstract

Norovirus (NV) is an infectious biological agent that causes gastrointestinal problems of the original nonbacterial appearance of foodborne illnesses. The genotype of NV responsible for the most frequent NV disease outbreaks is GII, accounting for 60–80% of the cases. Moreover, original and new NV variants are continuously emerging, concurrent with the recent global increase in NV infections. Hepatitis A virus (HAV) is another foodborne pathogen frequently implicated in acute gastroenteritis cases around the world. The virus is transmitted among humans via the fecal-oral route, and infection by HAV causes the most severe form of viral illness acquired from foods. In this study, we implemented primer sets to detect NV genotypes I and II. We also developed primer sets for the detection of HAV. The primers were used in a heminested reverse transcriptase PCR (hnRT-PCR) protocol that was rapid and sensitive for detecting NVG1, NVGII and HAV virus in food. The hnRT-PCR was applied successfully to strawberries and spinach obtained from a local fresh-food market, where we could see NVGI, NVGII and HAV. Keywords: Norovirus1; Hepatitis A2; gastroenteritis3; genotypes4; NVG15; NVGII6; hnRT-PCR7.